Compositions and methods useful in treatment of lower urinary tract symptoms, benign prostatic hyperplasia, erectile dysfunction

ABSTRACT

The invention relates to therapeutic compositions, solid oral dosage forms, and methods for treating, preventing, or alleviating lower urinary tract symptoms (LUTS), benign prostatic hyperplasia (BPH), erectile dysfunction (ED), urinary incontinence, bladder obstruction, overactive bladder (OAB), underactive bladder, interstitial cystitis, prostatitis, bladder and prostate inflammation, prostate fibrosis or pelvic pain. The therapeutic compositions can comprise cranberry powder enriched in cranberry seeds or cranberry seed meal.

This application is a continuation of International Application No.PCT/US2014/064961, filed Nov. 11, 2014, which claims the benefit ofpriority to U.S. Provisional Patent Application No. 61/902,376, filedNov. 11, 2013, the entire disclosures of which are incorporated byreference herein in their entirety.

2. FIELD

The present invention relates generally to treatment, prevention, andalleviation of lower urinary tract symptoms (LUTS), benign prostatichyperplasia (BPH), erectile dysfunction (ED), urinary incontinence, andother diseases or symptoms.

3. BACKGROUND

Lower urinary tract symptoms (LUTS) become increasingly bothersome asmen age, with a prevalence of moderate-to-severe symptoms rising tonearly 50% of men in their eighties. LUTS may or may not be related tobenign prostatic hyperplasia (BPH), a histological conditioncharacterized by the non-malignant overgrowth of prostatic tissuesurrounding the urethra that occurs in 50% of men in their fifties and90% of men in their eighties. LUTS can also arise from age-relatedbladder detrusor dysfunction and other sympathetic conditions. LUTS arefurther classified as voiding or storage symptoms and defined by theinternational prostate symptoms score (IPSS), a validated tool, widelyused among the medical and scientific community.

Voiding symptoms include urinary hesitancy, delay in initiatingmicturition, intermittency, involuntary interruption of voiding, weakurinary stream, straining to void, a sensation of incomplete emptying,terminal dribbling, and may be caused by prostate enlargement or tissueinflammation. Storage symptoms can include urinary frequency, nocturia,urgency, incontinence and bladder pain or dysuria, and may be caused bybladder detrusor overactivity. Physiological markers associated withincreased risk of BPH include high levels of testosterone,dihydrotestosterone, dehydroepiandrosterone and estradiol, insulin-likegrowth factors and inflammatory markers.

Although LUTS and LUTS due to BPH are not a life-threatening condition,the impact of LUTS on quality-of-life (QoL) can be significant andtreatment is necessary in most cases to avoid complications. Riskfactors include age, prostatic volume and peak urinary flow rate as wellas lifestyle, dietary pattern, alcohol consumption, physical activity orgenetic factors. Upon diagnosis, watchful waiting is recommended inapproximately 34% of cases in the United States. Registeredpharmacological treatments for LUTS may be responsible for a variety ofside effects, thus necessitating development of new treatments.

According to the American Urological Association, patients with a milddegree of bother (IPSS<8) or patients with moderate to severe symptoms(IPSS>8) who are not bothered by their LUTS may be managed with watchfulwaiting and lifestyle modification. If significant bothersome symptomspersist despite conservative measures, the initiation of medicalmanagement is indicated and in certain cases, surgery may berecommended.

The National Institutes of Health estimates that erectile dysfunction(ED) affects as many as 30 million men in the United States. Incidenceincreases with age: about 4% of men in their 50s and nearly 17% of menin their 60s experience a total inability to achieve an erection. Theincidence jumps to 47% for men older than 75.

Benign prostatic hyperplasia (BPH) is a common problem among older men,and is responsible for considerable disability. The prevalence ofhistologically diagnosed prostatic hyperplasia increases from 8% in menaged 31 to 40, to 40 to 50% in men aged 51 to 60, to over 80% in menolder than age 80.

Urinary incontinence is an underdiagnosed and underreported problem thatincreases with age. It affects 50-84% of the elderly in long-term carefacilities. At any age is more than 2 times more common in females thanin males.

Overactive bladder (OAB) is prevalent in 10 to 18% of the population,affecting men and women nearly equally. OAB has a negative impact onpatient's quality of life. LUTS associated with OAB are responsible ofsignificant social, psychological, occupational, domestic, and physicalstigmas.

4. SUMMARY

The invention is based, in part, on the discovery that addition ofcranberry seeds or cranberry seed meal to dried cranberry powderprovides a therapeutic composition that is effective against lowerurinary tract symptoms (LUTS) and other diseases or symptoms without theside effects of existing treatments.

In one aspect, the invention provides a therapeutic compositioncomprising dried cranberry powder and dried cranberry seeds.

In another aspect, the invention provides therapeutic compositioncomprising dried cranberry powder and cranberry seed meal.

In some embodiments, the cranberry is Vaccinium macrocarpon.

In some embodiments, the cranberry is Vaccinium microcarpon.

In some embodiments, the cranberry is Vaccinium oxycoccus.

In some embodiments, the cranberry seeds are present in an amount ofabout 5% to about 50% by weight of the dried cranberry powder.

In some embodiments, the cranberry seeds are present in an amount ofabout 15% to about 25% by weight of the dried cranberry powder.

In some embodiments, the cranberry seeds are present in an amount ofabout 20% by weight of the dried cranberry powder.

In some embodiments, the cranberry seed meal is present in an amount ofabout 5% to about 50% by weight of the dried cranberry powder.

In some embodiments, the cranberry seed meal is present in an amount ofabout 15% to about 25% by weight of the dried cranberry powder.

In some embodiments, the cranberry seed meal is present in an amount ofabout 20% by weight of the dried cranberry powder.

In some embodiments, the composition comprises less than about 12% oforganic acids by weight.

In some embodiments, the composition comprises less than 10% of organicacids by weight.

In some embodiments, the composition comprises about 5% to about 8% oforganic acids by weight.

In some embodiments, the composition comprises less than about 15% ofsugars by weight.

In some embodiments, the composition comprises less than about 12% ofsugars by weight.

In some embodiments, the composition comprises from about 1% to about 5%of quinic acid by weight.

In some embodiments, the composition comprises from about 2.2% to about3.2% of quinic acid by weight.

In some embodiments, the composition comprises from about 0.4% to about4% of malic acid by weight.

In some embodiments, the composition comprises from about 0.8% to about1.8% of malic acid by weight.

In some embodiments, the composition comprises from about 1% to about 5%of citric acid by weight.

In some embodiments, composition comprises from about 1.8% to about 3.2%of citric acid by weight.

In some embodiments, the composition comprises: 0.5% to 5.0%proanthocyanidins, 0.05% to 1.5% quercetin, 0.001% to 0.1%quercetin-3-glucoside, 0.001% to 0.1% quercetin-3-rhamnoside, 0.001% to0.1% quercetin-3-xyloside, 0.001% to 0.1% quercetin-3-arabinoside,0.001% to 0.5% myricetin, 0.001% to 0.1% peonidin-3-galactoside, 0.001%to 0.1% peonidin-3-glucoside, 0.001% to 0.1% peonidin-3-arabinoside,0.001% to 0.1% cyanidin-3-glucoside, 0.001% to 0.1%cyanidin-3-galactoside, 0.001% to 0.1% cyanidin-3-arabinoside, 0.001% to0.1% protocatechuic acid 0.001% to 0.1% p-coumaric acid, 0.001% to 0.1%caffeoyl-glucoside, 0.001% to 0.1% coumaroyl-glucoside, 0.001% to 0.1%cafeic acid, 0.001% to 0.1% chlorogenic acid or 0.01 to 1.5% ursolicacid by weight.

In some embodiments, the composition comprises: 1.0% to 1.2%proanthocyanidins, 0.16% to 0.20% quercetin, 0.07% to 0.09%quercetin-3-glucoside, 0.03% to 0.04% quercetin-3-rhamnoside, 0.019% to0.025% quercetin-3-xyloside, 0.025% to 0.035% quercetin-3-arabinoside,0.010% to 0.014% myricetin, 0.022% to 0.030% peonidin-3-galactoside,0.0025% to 0.0035% peonidin-3-glucoside, 0.010% to 0.020%peonidin-3-arabinoside, 0.0005% to 0.0015% cyanidin-3-glucoside, 0.015%to 0.030% cyanidin-3-galactoside, 0.010% to 0.025%cyanidin-3-arabinoside, 0.019% to 0.025% protocatechuic acid, 0.04% to0.06% p-coumaric acid, 0.015% to 0.025% caffeoyl-glucoside, 0.005% to0.015% coumaroyl-glucoside, 0.010% to 0.015% cafeic acid or 0.030% to0.04% chlorogenic acid by weight.

In some embodiments, the composition comprises about: 1.1%proanthocyanidins, 0.18% quercetin, 0.083% quercetin-3-glucoside, 0.034%quercetin-3-rhamnoside, 0.022% quercetin-3-xyloside, 0.030%quercetin-3-arabinoside, 0.012% myricetin, 0.027%peonidin-3-galactoside, 0.003% peonidin-3-glucoside, 0.014%peonidin-3-arabinoside, 0.001% cyanidin-3-glucoside, 0.022%cyanidin-3-galactoside, 0.018% cyanidin-3-arabinoside, 0.022%protocatechuic acid, 0.052% p-coumaric acid, 0.021% caffeoyl-glucoside,0.011% coumaroyl-glucoside, 0.014% cafeic acid, 0.034% chlorogenic acidor 0.92% ursolic acid by weight.

In some embodiments, the composition comprises 1 to 100 μglariciresinol, 1 to 100 μg secoisolariciresinol or 1 to 100μg/pinoresinol per 100 g of the composition by weight.

In some embodiments, the composition comprises about 51 μglariciresinol, about 12 μg secoisolariciresinol or about 78μg/pinoresinol per 100 g of the composition by weight.

In another aspect, the invention provides a solid oral dosage formcomprising a therapeutic composition described above.

In some embodiments, the solid oral dosage is a tablet.

In other embodiments, the solid oral dosage is a capsule.

In some embodiments, the solid oral dosage is a softgel.

In some embodiments, the solid oral dosage comprises 100 mg to 500 mg ofthe therapeutic composition.

In some embodiments, the solid oral dosage comprises 250 mg of thetherapeutic composition.

In some embodiments, the solid oral dosage comprises 500 mg of thetherapeutic composition.

In another aspect, the invention provides a method for alleviating lowerurinary tract symptoms (LUTS) in a subject comprising administering tothe subject in need thereof an effective amount of the compositiondescribed above.

In another aspect, the invention provides a method for alleviating thesymptoms of benign prostatic hyperplasia (BPH) in a subject comprisingadministering to the subject in need thereof an effective amount of thecomposition described above.

In another aspect, the invention provides a method for treating erectiledysfunction (ED) in a subject comprising administering to the subject inneed thereof an effective amount of the composition described above.

In another aspect, the invention provides a method for treating urinaryincontinence in a subject comprising administering to the subject inneed thereof an effective amount of the composition described above.

In another aspect, the invention provides a method for treatingoveractive bladder (OAB) in a subject comprising administering to thesubject in need thereof an effective amount of the composition describedabove.

In other aspects, the invention provides a method for treating bladderobstruction, interstitial cystitis, underactive bladder, prostatitis,bladder and prostate inflammation, prostate fibrosis or pelvic pain in asubject comprising administering to the subject in need thereof aneffective amount of the composition described above.

In some embodiments, the subject is a human

In some embodiments, the human is a male.

In some embodiments, the human is a female.

5. BRIEF DESCRIPTION OF THE DRAWINGS

The invention is described with reference to the following figures,which are presented for purposes of illustration only and which is notintending to be limiting of the invention.

FIG. 1 illustrates a CONSORT diagram of the 148 men attending the firstscreening visit in the LUTS study.

FIG. 2 illustrates a plot of the mean difference and corresponding 95%confidence interval in international prostate symptom scores (IPSS) atstart, 3-months, and 6-months for the placebo group, the therapeuticcomposition 250 mg group, and the therapeutic composition 500 mg groupin the LUTS study.

6. DETAILED DESCRIPTION 6.1 Definitions

The patent and scientific literature referred to herein establishesknowledge that is available to those of skill in the art. The issuedU.S. patents, allowed applications, published foreign applications, andreferences that are cited herein are hereby incorporated by reference tothe same extent as if each was specifically and individually indicatedto be incorporated by reference.

As used herein, the recitation of a numerical range for a variable isintended to convey that the invention may be practiced with the variableequal to any of the values within that range. Thus, for a variable whichis inherently discrete, the variable can be equal to any integer valuewithin the numerical range, including the end-points of the range.Similarly, for a variable which is inherently continuous, the variablecan be equal to any real value within the numerical range, including theend-points of the range. As an example, and without limitation, avariable which is described as having values between 0 and 2 can takethe values 0, 1 or 2 if the variable is inherently discrete, and cantake the values 0.0, 0.1, 0.01, 0.001, or any other real values >0 and<2 if the variable is inherently continuous.

As used herein, unless specifically indicated otherwise, the word “or”is used in the inclusive sense of “and/or” and not the exclusive senseof “either/or.”

As used herein, “about” means within ±10%. For example, “about 1” means“0.9 to 1.1”, “about 2%” means “1.8% to 2.2%”, “about 2% to 3%” means“1.8% to 3.3%”, and “about 3% to about 4%” means “2.7% to 4.4%.”

6.2 Cranberry Fruit

Cranberry fruit, e.g., Vaccinium macrocarpon, is recognized as a richsource of organic and phenolic acids, flavonols, flavan-3-ols,anthocyanins, proanthocyanidins (PACs) and pentacyclic triterpenoids,including ursolic and oleanolic acids.

A non-exhaustive list of pharmacoactive compounds that may be present incranberry fruit is listed in Table 1.

TABLE 1 Phenolic Coumaroyl-hexose Acid 6-Caffeoyl-D-glucose Deriva-3-caffeoylquinic acid (Chlorogenic acid) tives 4-hydroxybenzoic acid3-hydroxybenzoic acid 5-p-hydroxybenzoic acid 6-p-hydroxybenzoic acidGallic acid Benzoic acid Cinnamic acid dihydroxybenzoic acido-hydroxycinnamic acid 4-hydroxy-3-methoxybenzoic acid (HVA) (vanillicacid) 3-hydroxyphenylacetic acid (3HPAA) 3-hydroxybenzoic acid (3HBA)3,4-dihydroxybenzoic acid (3,4DHBA) (protocatechuic acid)2,3-dihydroxybenzoic acid (2,3DHBA) (hypogallic acid)2,4-dihydroxybenzoic acid (2,4DHBA) (b-resorcyclic acid)2,5-dihydroxybenzoic acid (2,5DHBA) (genistic acid) 2,6-dihydroxybenzoicacid 3-hydroxycinnamic acid (3HCA) 4-hydroxycinnamic acid (4HCA)(p-coumaric acid) 4-hydroxy-3-methoxycinnamic acid (Ferulic acid)3,4-dihydroxycinnamic acid (Caffeic acid) 3-hydroxyphenylpropionic acid(3HPPA) (phloretic acid) benzoylaminoacetic acid (hippuric acid)Salicylic acid Sinapic acid Ascorbic acid Ellagic acid Syringic acidShikimic acid Ferruloyl glucoside 4-caffeoylquinic acid(Cryptochlorogenic acid) Flavonols Myricetin Quercetin Quercetin3-galactoside Quercetin 3-glucoside Quercetin-3-rhamnosideQuercetin-3-xyloside Quercetin-3-arabinoside Quercetin glucuronideMyricetin-3-galactoside Myricetin-3-glucoside Isorhamnetin Flavanols(+)-catechin (−)-catechin Procyanidin A2 type isomer 1 Procyanidin A2type isomer 2 Procyanidin A2 type isomer 3 (−)-epicatechin3-methylcatechin 4-methylcatechin 3-methylepicatechin4-methylepicatechin Coumarins Scopoletin Antho- Cyanidin 3-galactosidecyanins Cyanidin 3-glucoside Peonidin 3-glucoside Cyanidin 3-arabinosidePeonidin 3-galactoside Peonidin 3-arabinoside Chalcones PhlorizinTocoph- Alpha-tocopherol acetate erols Alpha-tocopherol Beta-tocopherolGamma-tocopherol Delta-tocopherol Alpha-tocotrienol Beta-tocotrienolGamma-tocotrienol Delta-tocotrienol Vitamin E Sterols Cholesterol Brassicasterol 24 methyl-cholesterol Campestanol CampesterolDelta-7-campesterol Delta-5,23 stigmastadienol ClerosterolBeta-sitosterol Sitostanol Delta-5-avenasterolDelta-5,24-stigmastadienol Delta-7-stigmasterol Delta-7-avenasterolFatty acids Myristic acid Palmitic acid Palmitoleic acid Margaric acidStearic acid Oleic acid Linoleic acid Linolenic acid Arachidic acidGondoic acid Behenic acid Erucic acid Lignoceric acid Nervonic acidLignans Syringaresinol Lariciresinol MedioresinolLariciresinol-sesquilignan Secoisolariciresinol PinoresinolSecoisolariciresinol-cyclolariciresinol 7-hydroxymatairesinolNortrachelogenin 7-oxo-matairesinol Matairesinol α-conidendrin

6.3 Therapeutic Compositions Based on Cranberry Fruit Components

Therapeutic compositions described herein can include one or morecomponents described below in the ranges provided below.

In some embodiments, therapeutic compositions described herein comprisedried cranberry powder and dried cranberry seeds. In other embodiments,the therapeutic compositions described herein comprise dried cranberrypowder and cranberry seed meal.

The cranberry species used to make the compositions can be Vacciniummacrocarpon, Vaccinium microcarpon, or Vaccinium oxycoccus. However,other species of cranberries can also be used to make the compositionsdescribed herein.

Cranberry seeds can be present in the therapeutic composition in anamount of about 5% to about 50%, about 15% to about 25%, or about 20% byweight of the dried cranberry powder. Cranberry seed meal can be presentin the therapeutic composition in an amount of about 5% to about 50%,about 15% to about 25%, or about 20% by weight of the dried cranberrypowder.

A therapeutic composition can comprise less than about 12%, less thanabout 10%, or about 5% to about 8% of organic acids by weight. Exemplaryorganic acids include, but are not limited to, quinic acid, malic acid,and citric acid.

The organic acid content can be determined using the followingmodification of protocol AOAC 986.13 entitled Quinic, Malic and CitricAcids in Cranberry Juice Cocktail and Apple Juice (available from AOACInternational, www.aoac.org). The analytical column is a C18 reversephase column with a 5 μm particle size, 25 cm×4.6 mm in tandem with andfollowed by C18 reverse phase cartridges, with a 5 μm particle size and10 cm long. Phosphate buffer, 0.2M KH₂PO₄, pH 2.4 is used. The flow rateis 0.80 mL/min, the analysis is performed at ambient temperature, andthe detection wavelength is 214 nm A dilution of the test agent can beperformed in the mobile phase. CAS# for the standard are as follows:CAS-77-92-9 (citric acid), CAS-6915-15-7 (malic acid), CAS-77-95-2(quinic acid). The formula used to calculate the concentration of eachorganic acid is as follows: (PA/PA′)×(V′/V)×C, where PA and PA′=peakarea of test solution and standard, respectively; V and V′=volume oftest solution and standard, respectively; and C=concentration ofstandard, %. Shikimic acid and ascorbic acids can also be detected withthis method.

A therapeutic composition can comprise from about 1% to about 5% or fromabout 2.2% to about 3.2% of quinic acid by weight.

A therapeutic composition can comprise from about 0.4% to about 4% orfrom about 0.8% to about 1.8% of malic acid by weight.

A therapeutic composition can comprise from about 1% to about 5% or fromabout 1.8% to about 3.2% of citric acid by weight.

A therapeutic composition can comprise less than about 15% or less thanabout 12% of sugars by weight. Fructose, glucose and sucrose areexamples of sugars typically found in cranberry. The sugar contentreflects the global sugar amount and individual breakdown was notprovided on these lots.

The sugar content can be determined by HPLC, using the followingmodification of protocol AOAC 977.20. The column used isμ-Bondapak/Carbohydrate (Waters Associates, No. 84038) with a guardcolumn or equivalent. The mobile phase consists of non-spectroacetonitrile diluted with water (83/17, v/v). The sugar standardsolution consists of fructose (CAS#57-48-7), glucose (CAS#50-99-7) andsucrose (CAS#57-50-1). The sample is diluted in water and filteredthrough a 45 μm filter. 10 μL of sample are injected to the column atroom temperature. The flow rate is 1.0 mL/min under isocratic conditionsfor 20 min A refractive index detector is used. The amount of glucose,fructose, and sucrose is calculated from integrator values or from peakheights as follows: Weight % sugar=100×(PH/PH′)×(V/V′)×(W′/W) where PHand PH′=peak heights (or integrator values) of sample and standard,respectively; V and V′=mL sample and standard (50 and 100) solutions,respectively; and W and W′=g sample (5.000) and standard, respectively.

In some embodiments, the therapeutic composition comprises: 0.5% to 5.0%proanthocyanidins, 0.05% to 1.5% quercetin, 0.001% to 0.1%quercetin-3-glucoside, 0.001% to 0.1% quercetin-3-rhamnoside, 0.001% to0.1% quercetin-3-xyloside, 0.001% to 0.1% quercetin-3-arabinoside,0.001% to 0.5% myricetin, 0.001% to 0.1% peonidin-3-galactoside, 0.001%to 0.1% peonidin-3-glucoside, 0.001% to 0.1% peonidin-3-arabinoside,0.001% to 0.1% cyanidin-3-glucoside, 0.001% to 0.1%cyanidin-3-galactoside, 0.001% to 0.1% cyanidin-3-arabinoside, 0.001% to0.1% protocatechuic acid, 0.001% to 0.1% p-coumaric acid, 0.001% to 0.1%caffeoyl-glucoside, 0.001% to 0.1% coumaroyl-glucoside, 0.001% to 0.1%cafeic acid, 0.001% to 0.1% chlorogenic acid or 0.01 to 1.5% ursolicacid by weight.

The content of proanthocyanidins and one or more other components can bedetermined by HPLC-fluorescence using a Develosil Diol column orequivalent as follows. The column compartment is maintained at 35° C.The solvents used are as follows: (A) 2% acetic acid in acetonitrile and(B) is 95:3:2 methanol/water/acetic acid. A linear gradient is used from0% to 40% B, in 35 min; 40% to 100% B, in 40 min; 100% isocratic B, in45 min; and 100% to 0% B, in 50 min.

The content of anthocyanins (peonidin-3-galactoside,peonidin-3-glucoside, peonidin-3-arabinoside, cyanidin-3-glucoside,cyanidin-3-galactoside, cyanidin-3-arabinoside) can be determined byHPLC-UV at a wavelength of 535 nm. The column is Synergi Hydro-RP orequivalent. The column compartment is maintained at room temperature.The mobile phase A consists of an aqueous 5% formic acid solution andmobile phase B of methanol. The gradient applied is 0-2 min, 5% B; 2-10min, 5-20% B; 10-15 min, 20% B; 15-30 min, 20-25% B; 30-35 min, 25% B;35-50 min, 25-33% B; 50-55 min, 33% B; 55-65 min, 33-36% B; 65-70 min,36-45% B; 70-75 min, 45-53% B; 75-80 min, 53-55% B; 80-84 min, 55-70% B;84-88 min, 70-5% B; 88-90 min, 5% B.

The content of certain other phenolics can be determined by UPLC-MS/MSusing the following method. An Acquity T3 column (Waters Associates) orequivalent is used and placed in the UPLC column compartment maintainedat a temperature of 30° C. The solvents used are 0.1% Formic acid (A)and acetonitrile (B). A linear gradient is applied from 5% B; 0-4.5 min,5-20% B; 4.5-6.45 min, isocratic 20% B; 6.45-13.5 min, 20-45% B;13.5-16.5 min 45-100% B; 16.5-19.5 min isocratic 100% B; 19.5-19.52 min100-5% B; 19.52-22.5 min. The detection is done by MS/MS and allstandard compounds are tuned individually.

In some embodiments, the therapeutic composition comprises: 1.0% to 1.2%proanthocyanidins, 0.16% to 0.20% quercetin, 0.07% to 0.09%quercetin-3-glucoside, 0.03% to 0.04% quercetin-3-rhamnoside, 0.019% to0.025% quercetin-3-xyloside, 0.025% to 0.035% quercetin-3-arabinoside,0.010% to 0.014% myricetin, 0.022% to 0.030% peonidin-3-galactoside,0.0025% to 0.0035% peonidin-3-glucoside, 0.010% to 0.020%peonidin-3-arabinoside, 0.0005% to 0.0015% cyanidin-3-glucoside, 0.015%to 0.030% cyanidin-3-galactoside, 0.010% to 0.025%cyanidin-3-arabinoside, 0.019% to 0.025% protocatechuic acid, 0.04% to0.06% p-coumaric acid, 0.015% to 0.025% caffeoyl-glucoside, 0.005% to0.015% coumaroyl-glucoside, 0.010% to 0.015% cafeic acid or 0.030% to0.04% chlorogenic acid by weight.

In some embodiments, the therapeutic composition comprises about: 1.1%proanthocyanidins, 0.18% quercetin, 0.083% quercetin-3-glucoside, 0.034%quercetin-3-rhamnoside, 0.022% quercetin-3-xyloside, 0.030%quercetin-3-arabinoside, 0.012% myricetin, 0.027%peonidin-3-galactoside, 0.003% peonidin-3-glucoside, 0.014%peonidin-3-arabinoside, 0.001% cyanidin-3-glucoside, 0.022%cyanidin-3-galactoside, 0.018% cyanidin-3-arabinoside, 0.022%protocatechuic acid, 0.052% p-coumaric acid, 0.021% caffeoyl-glucoside,0.011% coumaroyl-glucoside, 0.014% cafeic acid, 0.034% chlorogenic acidor 0.92% ursolic acid by weight.

In some embodiments, the therapeutic composition comprises 1 to 100 μgor about 51 lariciresinol per 100 g of the composition. In someembodiments, the therapeutic composition comprises 1 to 100 μg or about12 μg secoisolariciresinol per 100 g of the composition. In someembodiments, the therapeutic composition comprises 1 to 100 μg or about78 μg pinoresinol per 100 g of the composition.

The content of lignans can be determined by UPLC-MS/MS using an ACQUITYBEH C18 reverse phase column or equivalent as follows. The columncompartment is maintained at 30° C. The solvents used are (A) Formicacid 0.1% and (B) Acetonitrile. The gradient is 5% B, 8.0 min, 30% B,9.0 min, 30% B, 10.0 min, 50% B, 12.0 min, 50% B, 15.0 min, 95% B, 170min 95% B, 17.5 min, 5% B, 23.0 min, 5% B. The detection is done byMS/MS and all standard compounds are tuned individually.

In some embodiments, the therapeutic compositions described above can beproduced without cranberry fruit, e.g., by combining ingredientsobtained from natural sources or by chemical synthesis into atherapeutic composition.

6.4 Solid Dosage Forms and Methods of Use

In some embodiments, a solid oral dosage comprising the therapeuticcomposition described herein is a tablet, a capsule, or a softgel. Insome embodiments, such solid oral dosage comprises from 50 mg to 500 mgof the therapeutic composition, e.g., 50 mg, 100 mg, 150 mg, 200 mg, 250mg, 300 mg, 350 mg, 400 mg, 450 mg or 500 mg.

Solid dosage forms comprising the therapeutic compositions describedherein optionally comprise a suitable amount of one or morepharmaceutically acceptable excipients so as to provide the form forproper administration to the subject.

Such pharmaceutical excipients can be liquids, such as water and oils,including those of petroleum, animal, vegetable, or synthetic origin,such as peanut oil, soybean oil, mineral oil, sesame oil and the like.The pharmaceutical excipients can be saline, gum acacia, gelatin, starchpaste, talc, keratin, colloidal silica, urea and the like. In addition,auxiliary, stabilizing, thickening, lubricating, and coloring agents canbe used. In one embodiment, the pharmaceutically acceptable excipientsare sterile when administered to a subject. Suitable pharmaceuticalexcipients also include starch, glucose, lactose, sucrose, gelatin,malt, rice, flour, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene glycol, water, ethanol and the like. The present therapeuticcompositions, if desired, can also contain minor amounts of wetting oremulsifying agents, or pH buffering agents. In one embodiment, thecomposition is in the form of a capsule (see, e.g., U.S. Pat. No.5,698,155). Other examples of suitable pharmaceutical excipients aredescribed in Remington's Pharmaceutical Sciences 1447-1676 (Alfonso R.Gennaro eds., 19th ed. 1995).

In some embodiments, a therapeutic composition described herein isformulated in accordance with routine procedures as a compositionadapted for oral administration to human beings. Compositions for oraldelivery can be in the form of tablets, lozenges, aqueous or oilysuspensions, granules, powders, emulsions, capsules, softgels, syrups,or elixirs for example. Orally administered compositions can contain oneor more agents, for example, sweetening agents such as fructose,aspartame or saccharin; flavoring agents such as peppermint, oil ofwintergreen, or cherry; coloring agents; and preserving agents, toprovide a pharmaceutically palatable preparation. Moreover, where intablet or pill form, the compositions can be coated to delaydisintegration and absorption in the gastrointestinal tract therebyproviding a sustained action over an extended period of time.Selectively permeable membranes surrounding an osmotically activetherapeutic composition is also suitable for orally administeredcompositions. In these latter platforms, fluid from the environmentsurrounding the capsule is imbibed by the driving compound, which swellsto displace the agent or agent composition through an aperture. Thesedelivery platforms can provide an essentially zero order deliveryprofile as opposed to the spiked profiles of immediate releaseformulations. A time delay material such as glycerol monostearate orglycerol stearate can also be useful. Oral compositions can includestandard excipients such as mannitol, lactose, starch, maltodextrin,cyclodextrins, alginate, arabic or guar gum, magnesium stearate, sodiumsaccharin, cellulose, and magnesium carbonate. In one embodiment, theexcipients are of pharmaceutical grade.

Pharmaceutical dosage forms for oral use can be obtained throughcombination of a therapeutic composition described herein with a solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable additional compounds, ifdesired, to obtain tablets or dragee cores. Suitable solid excipients inaddition to those previously mentioned are carbohydrate or proteinfillers that include, but are not limited to, sugars, including lactose,sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato,or other plants; cellulose such as methyl cellulose,hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; and gumsincluding arabic and tragacanth; as well as proteins such as gelatin andcollagen. Maltodextrin and cyclodextrins can also be used. If desired,disintegrating or solubilizing agents may be added, such as thecross-linked polyvinyl pyrrolidone, agar, alginic acid, or a saltthereof, such as sodium alginate.

Capsules for oral use include hard gelatin capsules in which the activeingredient is mixed with a solid diluent, and soft gelatin capsuleswherein the active ingredients is mixed with water or an oil such aspeanut oil, liquid paraffin or olive oil.

Softgels for oral use may consist of a gelatin based shell surrounding aliquid fill. Softgel shells can be made of a combination of gelatin,water, opacifier and a plasticiser such as glycerin and/or sorbitol.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Therapeutic compositions described herein can be administered bycontrolled-release or sustained release means or by delivery devicesthat are well known to those of ordinary skill in the art. Examplesinclude, but are not limited to, those described in U.S. Pat. Nos.5,674,533; 5,059,595; 5,120,548; 5,073,543; 5,639,476 and 5,354,556,each of which is incorporated herein by reference in its entirety. Suchdosage forms can be useful for providing controlled or sustained releaseof one or more active ingredients using, for example, hydropropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmoticsystems, multilayer coatings, microparticles, liposomes, microspheres,or a combination thereof to provide the desired release profile invarying proportions. Suitable controlled or sustained releaseformulations known to those skilled in the art, including thosedescribed herein, can be readily selected for use with the activeingredients of the invention. The invention thus encompasses single unitdosage forms suitable for oral administration such as, but not limitedto, tablets, capsules, gelcaps, and caplets that are adapted forcontrolled or sustained release.

In some embodiments, a controlled or sustained release compositioncomprises a minimal amount of a therapeutic composition to alleviate thesymptoms of, treat or prevent lower urinary tract symptoms (LUTS),benign prostatic hyperplasia (BPH), erectile dysfunction (ED), urinaryincontinence, bladder obstruction, interstitial cystitis, overactivebladder (OAB), underactive bladder, prostatitis, bladder and prostateinflammation, prostate fibrosis or pelvic pain in a patient over aperiod of time. Advantages of controlled or sustained releasecompositions include extended activity of the drug, reduced dosagefrequency, and increased subject compliance. In addition, controlled orsustained release compositions can favorably affect the time of onset ofaction or other characteristics, such as blood levels active ingredientspresent in the therapeutic composition, and can thus reduce theoccurrence of adverse side effects.

Controlled or sustained release compositions can initially release anamount of an active ingredient present in the therapeutic compositionthat promptly produces the desired therapeutic or prophylactic effect,and gradually and continually release other amounts of the activeingredients present in the therapeutic composition to maintain thislevel of therapeutic or prophylactic effect over an extended period oftime. To maintain a constant level of an active ingredient present inthe therapeutic composition in the body, active ingredients present inthe therapeutic composition thereof can be released from the dosage format a rate that will replace the amount of the active ingredients presentin the therapeutic composition being metabolized and excreted from thebody. Controlled or sustained release of an active ingredient can bestimulated by various conditions, including but not limited to, changesin pH, changes in temperature, concentration or availability of enzymes,concentration or availability of water, or other physiologicalconditions or compounds.

The amount of a therapeutic composition that is effective in alleviatingthe symptoms of, treating or preventing lower urinary tract symptoms(LUTS), benign prostatic hyperplasia (BPH), erectile dysfunction (ED),urinary incontinence, bladder obstruction, interstitial cystitis,overactive bladder (OAB), underactive bladder, prostatitis, bladder andprostate inflammation, prostate fibrosis or pelvic pain can bedetermined by standard clinical techniques. In addition, in vitro or invivo assays can optionally be employed to help identify optimal dosageranges. The precise dose to be employed can also depend on the route ofadministration, and the seriousness of the condition being treated andcan be decided according to the judgment of the practitioner and eachsubject's circumstances in view of, e.g., published clinical studies.Suitable effective dosage amounts, however, range from about 1 mg toabout 5 grams about every 24 hours, although they are typically about500 mg or less per every 24 hours. In one embodiment, the effectivedosage is about 50 mg, about 100 mg, about 150 mg, about 200 mg, about250 mg, about 300 mg, about 350 mg about 400 mg, about 450 mg about 500mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1 g,about 1.2 g, about 1.4 g, about 1.6 g, about 1.8 g, about 2.0 g, about2.2 g, about 2.4 g, about 2.6 g, about 2.8 g, about 3.0 g, about 3.2 g,about 3.4 g, about 3.6 g, about 3.8 g, about 4.0 g, about 4.2 g, about4.4 g, about 4.6 g, about 4.8 g, and about 5.0 g, every 24 hours.Equivalent dosages can be administered over various time periodsincluding, but not limited to, about every 2 hours, about every 4 hours,about every 6 hours, about every 8 hours, about every 24 hours, aboutevery 36 hours, about every 48 hours, about every 72 hours, about everyweek, about every two weeks, about every three weeks, about every month,and about every two months. The effective dosage amounts describedherein refer to total amounts administered; that is, if more than onetherapeutic composition is administered, the effective dosage amountscorrespond to the total amount administered.

The therapeutic composition can be administered as long as the symptomspersist or longer. In some embodiments, the therapeutic composition isadministered for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days.

The dosage regimen utilizing the therapeutic compositions describedherein can be selected in accordance with a variety of factors includingtype, species, age, weight, sex and medical condition of the subject;the severity of the condition to be treated; the route ofadministration; and the renal or hepatic function of the subject.

A therapeutic composition described herein can be administered in asingle daily dose, or the total daily dosage can be administered individed doses of two, three or four times daily.

The therapeutic composition described herein can be assayed in vitro orin vivo for the desired therapeutic or prophylactic activity prior touse in humans. Animal model systems can be used to demonstrate safetyand efficacy.

5. Kits

Described herein are kits that can simplify the administration of atherapeutic composition described herein to a subject. A typical kitcomprises a unit dosage form of a therapeutic composition describedherein and a label or printed instructions. In some embodiments, thelabel or printed instructions instruct the use of the unit dosage formto alleviate the symptoms of, treat or prevent lower urinary tractsymptoms (LUTS), benign prostatic hyperplasia (BPH), erectiledysfunction (ED), urinary incontinence, bladder obstruction,interstitial cystitis, overactive bladder (OAB), underactive bladder,prostatitis, bladder and prostate inflammation, prostate fibrosis orpelvic pain.

The kit can also further comprise a unit dosage form of anotherprophylactic or therapeutic agent. Examples of other prophylactic ortherapeutic agents include, but are not limited to, those listed above.

6.4 Methods Of Making Therapeutic Compositions

Therapeutic compositions described herein can be made as follows.Cranberry berries, skins, juice or pomace are weighed and mixed tocreate a slurry. If the composition is enriched in cranberry seeds,seeds are added to the slurry. Water is added to the slurry to reachabout 5%-15% of the solids by weight. The slurry is passed through a 14mesh screen, a shear pump, spray-dried and milled. If the therapeuticcomposition is enriched in cranberry seed meal, it is added during themilling. The milled composition is passed through a sieve, sealed andpelletized.

6.5 Methods Of Using Therapeutic Compositions 6.5.1 Use in the Treatmentof Lower Urinary Tract Symptoms (LUTS)

Therapeutic compositions described herein are useful for alleviating thesymptoms of, treating or preventing lower urinary tract symptoms (LUTS).Accordingly, described herein are methods for alleviating, treating orpreventing lower urinary tract symptoms (LUTS) in a subject comprisingadministering to the subject in need thereof an effective amount of atherapeutic composition described herein. LUTS may or may not be relatedto benign prostatic hyperplasia (BPH), a histological conditioncharacterized by the non-malignant overgrowth of prostatic tissuesurrounding the urethra. The alleviation of symptoms can be manifestedby comparison to the same subject prior to administration or treatmentwith the therapeutic composition. The subject can be a human, forexample a male or a female. In some embodiments, the human is older than45 years, older than 50 years, older than 55 years, older than 60 years,older than 65 years, older than 70 years, older than 75 years or olderthan 80 years.

6.5.2 Use in the Treatment of Benign Prostatic Hyperplasia (BPH)

Benign prostatic hyperplasia (BPH), a histological conditioncharacterized by the non-malignant overgrowth of prostatic tissuesurrounding the urethra.

The therapeutic compositions described herein are useful for alleviatingthe symptoms of, treating or preventing to benign prostatic hyperplasia(BPH). Accordingly, described herein are methods for alleviating thesymptoms of, treating or preventing benign prostatic hyperplasia (BPH)in a subject comprising administering to the subject in need thereof aneffective amount of a therapeutic composition described herein. Thealleviation of symptoms of benign prostatic hyperplasia (BPH) can bemanifested by comparison to the same subject prior to administration ortreatment with the therapeutic composition. The subject can be a human,for example a male. In some embodiments, the human is older than 45years, older than 50 years, older than 55 years, older than 60 years,older than 65 years, older than 70 years, older than 75 years or olderthan 80 years.

6.5.3 Use in the Treatment of Erectile Dysfunction (ED)

The therapeutic compositions described herein are useful for alleviatingthe symptoms of, treating or preventing benign erectile dysfunction(ED). Accordingly, described herein are methods for alleviating thesymptoms of, treating or preventing erectile dysfunction (ED) in asubject comprising administering to the subject in need thereof aneffective amount of a therapeutic composition described herein. Thealleviation of symptoms of erectile dysfunction (ED) can be manifestedby comparison to the same subject prior to administration or treatmentwith the therapeutic composition. The subject can be a human, forexample a male. In some embodiments, the human is older than 45 years,older than 50 years, older than 55 years, older than 60 years, olderthan 65 years, older than 70 years, older than 75 years or older than 80years.

6.5.4 Use in the Treatment of Urinary Incontinence

The therapeutic compositions described herein are useful for alleviatingthe symptoms of, treating or preventing urinary incontinence.Accordingly, described herein are methods for alleviating the symptomsof, treating or preventing urinary incontinence in a subject comprisingadministering to the subject in need thereof an effective amount of atherapeutic composition described herein. The alleviation of symptoms ofurinary incontinence can be manifested by comparison to the same subjectprior to administration or treatment with the therapeutic composition.The subject can be a human, for example a male or a female. In someembodiments, the human is older than 45 years, older than 50 years,older than 55 years, older than 60 years, older than 65 years, olderthan 70 years, older than 75 years or older than 80 years.

6.5.5 Use in the Treatment of Overactive Bladder (OAB)

The therapeutic compositions described herein are useful for alleviatingthe symptoms of, treating or preventing overactive bladder (OAB).Accordingly, described herein are methods for alleviating the symptomsof, treating or preventing overactive bladder in a subject comprisingadministering to the subject in need thereof an effective amount of atherapeutic composition described herein. The alleviation of symptoms ofoveractive bladder (OAB) can be manifested by comparison to the samesubject prior to administration or treatment with the therapeuticcomposition. The subject can be a human, for example a male or a female.In some embodiments, the human is older than 45 years, 50 years, olderthan 55 years, older than 60 years, older than 65 years, older than 70years, older than 75 years or older than 80 years.

6.5.6 Use in the Treatment of Additional Diseases or Symptoms

The therapeutic compositions described herein are useful for alleviatingthe symptoms of, treating or preventing bladder obstruction,interstitial cystitis, underactive bladder, prostatitis, bladder andprostate inflammation, prostate fibrosis or pelvic pain. The alleviationof symptoms of these diseases or conditions can be manifested bycomparison to the same subject prior to administration or treatment withthe therapeutic composition. The subject can be a human, for example amale or a female. In some embodiments, the human is older than 45 years,older than 50 years, older than 55 years, older than 60 years, olderthan 65 years, older than 70 years, older than 75 years or older than 80years.

7. EXAMPLES

This invention is further illustrated by the following examples, whichshould not be construed as limiting. Those skilled in the art willrecognize, or be able to ascertain, using no more than routineexperimentation, numerous equivalents to the specific substances andprocedures described herein. Such equivalents are intended to beencompassed in the scope of the claims that follow the examples below.

7.1 Example 1 Preparation of Therapeutic Composition Using CranberrySeed Meal

Therapeutic compositions described herein can be made as follows.Cranberry berries, skins, juice or pomace are weighed and mixed tocreate a slurry. If the composition is enriched in cranberry seeds,seeds are added to the slurry. Water is added to the slurry to reachabout 5%-15% of the solids by weight. The slurry is passed through a 14mesh screen, a shear pump, spray-dried and milled. If the therapeuticcomposition is enriched in cranberry seed meal, it is added during themilling. The milled composition is passed through a sieve, sealed andpelletized.

7.2 Example 2 Characterization of the Therapeutic Composition

The therapeutic compositions were prepared as described above andcharacterized. Table 2 shows the organic acid profile of 5 samples asdetermined using a modified AOAC (986.13) entitled Quinic, Malic andCitric Acids in Cranberry Juice Cocktail and Apple Juice. Analyticalcolumn is a C18 reverse phase column with a 5 μm particle size, 25cm×4.6 mm in tandem with and followed by C18 reverse phase cartridges,with a 5 μm particle size and 10 cm long. Phosphate buffer, 0.2M KH₂PO₄,pH 2.4 is used. The elution is isocratic, flow rate 0.80 mL/min, ambienttemperature and wavelength 214 nm. The method was originally written forcranberry juice and adapted for powders, where a dilution of the testagent was performed in the mobile phase. CAS# for the standard are asfollow: CAS-77-92-9 (citric acid), CAS-6915-15-7 (malic acid),CAS-77-95-2 (quinic acid). The formula used to calculate theconcentration of each organic acid is as follow: (PA/PA′)×(V′/V)×C,where PA and PA′=peak area of test solution and standard, respectively;V and V′=volume of test solution and standard, respectively; andC=concentration of standard, %. The results are presented in Table 2below.

TABLE 2 Quinic Malic Sample Acid (%) Acid (%) Citric Acid (%) Total (%)1 2.43 1.54 2.38 6.35 2 2.31 0.93 1.94 5.18 3 2.99 1.43 2.99 7.41 4 2.401.36 2.15 5.91 5 2.75 1.65 3.00 7.40 Mean ± SD 2.58 ± 0.28 1.38 ± 0.282.49 ± 0.48 6.45 ± 0.97

Table 3 shows the sugar content of 5 samples as determined by HPLC,using a modification of AOAC method AOAC 977.20. The column used is a300×4 (id) mm μ-Bondapak/Carbohydrate (Waters Associates, No. 84038)with a guard column. The mobile phase consists of nonspectroacetonitrile diluted with water (83/17, v/v). The sugar standardsolution consisted of fructose (CAS#57-48-7), glucose (CAS#50-99-7) andsucrose (CAS#57-50-1). The sample was diluted in water and filteredthrough a 45 μm filter. 10 μL of sample are injected to the column atroom temperature. The flow rate is 1.0 mL/min under isocratic conditionsfor 20 min A refractive index detector is used. Glucose, fructose, andsucrose are calculated from integrator values or from peak heights asfollows: Weight % sugar=100×(PH/PH′)×(V/V′)×(W′/W) where PH and PH′=peakheights (or integrator values) of sample and standard, respectively; Vand V′=mL sample and standard (50 and 100) solutions, respectively; andW and W′=g sample (5.000) and standard, respectively. The results arepresented in Table 3 below.

TABLE 3 Sample Sugar Content (g/100 g) 6 8.7 7 4.0 8 7.7 9 5.9 10  7.3Mean ± SD 6.7 ± 1.8

Concentration of proanthocyanidins was analyzed by HPLC-fluorescenceusing a Develosil Diol or equivalent. The column size is 250 mm×4.6 mmwith a 5 micron particle size. The column compartment is maintained at35° C. The solvents used are as follows: (A) 2% acetic acid inacetonitrile and (B) is 95:3:2 methanol/water/acetic acid. A lineargradient was used from 0% to 40% B, in 35 min; 40% to 100% B, in 40 min;100% isocratic B, in 45 min; and 100% to 0% B, in 50 min

The anthocyanins (peonidin-3-galactoside, peonidin-3-glucoside,peonidin-3-arabinoside, cyanidin-3-glucoside, cyanidin-3-galactoside,cyanidin-3-arabinoside) are analyzed by HPLC-UV at a wavelength of 535nm. The column is Synergi Hydro-RP 250 mm×4, 6 mm with a 4 μm particlesize. The column compartment is maintained at room temperature. Themobile phase A consists of an aqueous 5% formic acid solution and mobilephase B of methanol. The gradient applied is 0-2 min, 5% B; 2-10 min,5-20% B; 10-15 min, 20% B; 15-30 min, 20-25% B; 30-35 min, 25% B; 35-50min, 25-33% B; 50-55 min, 33% B; 55-65 min, 33-36% B; 65-70 min, 36-45%B; 70-75 min, 45-53% B; 75-80 min, 53-55% B; 80-84 min, 55-70% B; 84-88min, 70-5% B; 88-90 min, 5% B.

The rest of the phenolics are analyzed by UPLC-MS/MS using the followingmethod. An Acquity T3 column (150 mm×2.1 mm id, 1.8 μm particle size)from Waters Associates is used and placed in the UPLC column compartmentmaintained at a temperature of 30° C. The solvents used are 0.1% Formicacid (A) and acetonitrile (B). A linear gradient is applied from 5% B;0-4.5 min, 5-20% B; 4.5-6.45 min, isocratic 20% B; 6.45-13.5 min, 20-45%B; 13.5-16.5 min 45-100% B; 16.5-19.5 min isocratic 100% B; 19.5-19.52min 100-5% B; 19.52-22.5 min. The detection was done by MS/MS and allstandard compounds were tuned individually. The results are shown inTable 4.

TABLE 4 Concentration Class Subclass Phytochemical (mg/100 g) Flavo-Flavan-3-ols Proanthocyanidins 1078.9* noids Flavonols Quercetin 176.6Quercetin-3-glucoside 83.2 Quercetin-3-rhamnoside 34.2Quercetin-3-xyloside 21.9 Quercetin-3-arabinoside 30.2 Myricetin 11.8Anthocyanin Peonidin-3-galactoside 27.2 Peonidin-3-glucoside 2.9Peonidin-3-arabinoside 14.3 Cyanidin-3-glucoside 1.1Cyanidin-3-galactoside 22.5 Cyanidin-3-arabinoside 17.7 PhenolicHydroxybenzoic Protocatechuic acid 21.7 acids Hydroxycinnamic p-Coumaricacid 51.7 acid Caffeoyl-glucoside 21.2 Coumaroyl-glucoside 10.7 Cafeicacid 14.3 Chlorogenic acid 33.8 *Measured by UPLC-MS/MS.

Proanthocyanidin levels are typically >0.3% when measured using theBL-DMAC method (Prior et al., “Multi-laboratory validation of a standardmethod for quantifying proanthocyanidins in cranberry powders” J SciFood Agric. 90:1473-8 (2010)), >1.5% when measured using a modified HPLCmethod in which catechin monomers (quantified by HPLC-UV) are subtractedfrom total polyphenols measured by folin (Sakakibara et al.,“Simultaneous Determination of All Polyphenols in Vegetables, Fruits andTeas” J. Agri. Food. Chem., Vol. 51. Pp 572-580 (2003); Methods inEnzymology, Volume 299, “Oxidants and Antioxidants Part A” Pages152-178, 1999 (modified)), or >5% when using a modified EuropeanPharmacopeia method (European Pharmacopoeia 6.0; 01/2008:1220), and >1%by HPLC-fluorescence.

Lignans are analyzed by UPLC-MS/MS using a 2.1 mm×150 mm ACQUITY BEH C18reverse phase 1.7 μm particle size column. The column compartment wasmaintained at 30° C. The solvents used are (A) Formic acid 0.1% and (B)Acetonitrile. The gradient is 5% B, 8.0 min, 30% B, 9.0 min, 30% B, 10.0min, 50% B, 12.0 min, 50% B, 15.0 min, 95% B, 17.0 min 95% B, 17.5 min,5% B, 23.0 min, 5% B. The detection was done by MS/MS and all standardcompounds were tuned individually. The results are shown in Table 5.

TABLE 5 Name Concentration (μg/100 g) Lariciresinol 51.3Lariciresinol-sesquilignan 3.8 Secoisolariciresinol 12.0 Pinoresinol77.8

7.3 Example 3 Determination of Efficacy of Therapeutic CompositionsAgainst LUTS

The therapeutic composition supplied by NATUREX-DBS LLC., USA, was used.Capsules consisted of either 500 mg of the therapeutic composition, or acombination of 250 mg of the therapeutic composition and 250 mg ofplacebo, or 500 mg of placebo. The composition of the placebo was asfollows: low density STAR-DRI® 1015A maltodextrin, canola oil, Red 40Lake, sodium aluminum silicate and Blue 1 Lake. The capsules wereindistinguishable in appearance. All capsules were provided in identicalplastic boxes with safe seal.

The study was a 6-month, single-center, randomized, double-blind,placebo controlled trial, consisting of three parallel treatment arms.Inclusion criteria comprised of: IPSS score between 8 to 19, individualsolder than 45 years old and prostate specific antigen (PSA) values lowerthan 2.5 ng/mL. Exclusion criteria included food allergies, recentprostatitis, chronic liver or kidney diseases, as well as neurological,gastrointestinal or metabolic disorder or any other chronic healthcondition such as diabetes, excess consumption of caffeine or alcohol.Subjects were also ineligible if they had prior invasive treatment forBPH, recent treatment with α-blockers (within 1 month) or 5α-reductaseinhibitors (within 6 months), phytotherapy including saw palmetto,beta-sitosterol, pygeum or other complementary therapy (within 3months). The primary outcome measure was the IPSS, evaluated atbaseline, 3 and 6 months. Secondary outcome measures included quality oflife (QoL) at baseline, 3 and 6 months, as well as bladder voided volume(Vol), maximum urinary flow rate (Qmax), average urinary flow rate(Qave) and ultrasound estimated post-void residual urine volume (PVR),serum PSA, selenium, interleukin-6 (IL-6) and C-reactive protein (CRP),at baseline and 6 months.

Participants were randomly assigned to consume daily 500 mg of thetherapeutic composition (n=40), 250 mg of the therapeutic composition(n=43) or placebo (n=41) for 6 months. The randomization plan fortreatment assignment to subjects was generated with on line softwareQuickCalcs (GraphPad Software Inc., USA, last accessed on Jul. 2, 2014)and carried out by study staff.

Participants were observed at baseline, on day-90 (3-month) and day-180(6-month). During the health examination on the first day, and day-180the following actions were required and health parameters were assessed:(i) detailed medical history, (ii) assessment of all concurrent medicaldrugs and therapies, (iii) dietary habits, (iv) completion of the IPSSquestionnaire, including a question on QoL, (v) urinanalysis, (vi)uroflowmetry, (vii) kidney and bladder ultrasound and (viii) a bloodlaboratory analysis including PSA. On day-90, only the physicalexamination and IPSS score were performed. The therapeutic compositionbottles were collected at day-90 and at the end of study. Compliance wasassessed by performing remaining capsule counts.

Uroflowmetry data: Qmax and Qave were measured using FlowMic(Medkonsult, Czech Republic). The subjects were instructed not tourinate for several hours before the test and to drink at least 1 L offluids to ensure a full bladder. The Qmax and Qave were calculated bymeasuring the Vol per unit of time. PVR was assessed using an ultrasounddevice BK Medical Viking 2400 with abdominal probe 3-7 MHz. Vol and PVRwere calculated using the formula for a prolate ellipsoid(width×length×height×0.523). Qmax, Qave, Vol and PVR were measured onday-0 and day-180.

Basic biochemical and hematological parameters were determined in allsamples using a HITACHI Modular Evo P analyzer (Hitachi, Japan). SerumPSA was determined using an Architect type LEIA analyzer (AbbottLaboratories, Abbott Park, Ill., USA). CRP was determined by a Quikread101 and IL-6 by the system Modular® Analytics. Selenium in plasma wasestimated by atomic absorption spectrometry using the AA6300 instrument(Shimadzu, Japan). Hemoglobin (Hb), hematocrit (Htc), erythrocytes(RBC), thrombocytes (PLT) and leukocytes (WBC) were measured in Na₂EDTAblood.

The data of the treatment groups was compared with respect to baselinemeasures using the Wilcoxon matched paired test. The primary andsecondary analyses were based on the per protocol population thatincluded all eligible participants who were treated during the entirelength of the study. A Mann-Whitney U test was used to compare bothtreatment dose to placebo data. P-values <0.05 were considered to besignificant.

An analysis of covariance was used to test whether there was an effectof the dose on the outcome measure at the end of treatment. Thisanalysis of covariance used the daily dose units of 250 mg as acontinuous variable and the baseline measurements of the outcome as acovariate. Box plots of residuals were examined against dose (0, 250,500 mg) to determine whether the residuals were bell-shaped and whetherthere was an indication of a non-linear dose effect.

PVR was considered to follow a two-stage process because some proportionof participants had no measurable PVR. This proportion was modelled as abinomial distribution. The volume of urine among participants with PVRwas modelled using a truncated Poisson distribution. This two stagemodel was fit using the hurdle function in the ‘pscl’ package (StanfordUniversity) running on R version 3.0.0. Dose/250 mg, baseline PVR andbaseline IPSS were entered into this model.

A total of 148 men were pre-screened for the study. FIG. 1 provides aCONSORT diagram of the 148 men attending the first screening visit. Atotal of 124 men were randomized, 41 to the placebo group, 43 to thetherapeutic composition 250 mg group and 40 to the therapeuticcomposition 500 mg group. In the therapeutic composition 500 mg group, 2participants were lost to follow-up and were not included in the perprotocol analysis.

Table 6 presents a summary of baseline characteristics and LUT functionmeasures across the three groups of the analysis. Results are presentedas mean±standard deviation (SD).

TABLE 6 Therapeutic Therapetic Composition Composition Placebo 250 mg500 mg (n = 41) (n = 43) (n = 38) Age (years) 54.0 ± 5.1 53.3 ± 5.2 52.5± 5.4 Weight (kg)  89.3 ± 11.9  91.2 ± 11.9 90.1 ± 8.0 Height (cm) 178.5± 6.6  180.6 ± 6.6  180.7 ± 6.2  Body Mass Index 28.1 ± 3.8 27.9 ± 2.927.7 ± 3.0 Systolic Blood 131.1 ± 12.1 130.6 ± 10.1 132.1 ± 11.5Pressure (mmHg) Diastolic Blood 80.1 ± 7.3 80.5 ± 7.2 80.9 ± 7.4Pressure (mmHg) Heart Beat (bpm) 68.9 ± 3.5 67.8 ± 4.6 68.1 ± 4.4 IPSS(score)  9.1 ± 2.0  9.7 ± 3.1  9.4 ± 2.0 PVR (mL)  15.0 ± 19.2  15.9 ±23.2  17.8 ± 21.0 Q_(max) (mL/sec) 22.0 ± 7.8 20.5 ± 7.1 19.5 ± 7.5Q_(ave) (mL/sec) 14.3 ± 5.2 12.5 ± 4.6 12.5 ± 5.5 Bladder Volume (mL) 408.5 ± 117.9  339.9 ± 114.4  339.0 ± 118.9

Participants had a mean age of 53.3±5.4 years with a mean IPSS score of9.4±2.4. Adherence with scheduled visits was 98.4%. Compliance to thetreatment was 100%. IPSS data with voiding and storage symptom subscoreand QoL data during the 6-month treatment period are presented in Table7. The voiding and storage subscores corresponded to questions 1(incomplete emptying), 3 (intermittency), 5 (weak stream), 6 (straining)and questions 2 (frequency), 4 (urgency), 7 (nocturia), respectively.There was a decrease in IPSS score in the groups taking the therapeuticcomposition at 250 mg and at 500 mg at the end of the 6-month periodwith a significant difference versus placebo (p=0.05 and p<0.001,respectively using a Mann-Whitney U test).

Participants IPSS score, voiding and storage symptom score and qualityof life score at baseline (Day-0), 3 months (Day-90) and 6 months(Day-180) after placebo, the therapeutic composition at 250 mg or 500 mgintake are shown in Table 7 and Table 8. In Table 7, results arepresented as mean±SD and as median along with first and third quartiles.

TABLE 7 Day-0 Day-90 Day-180 % change at Group Mean ± SD Median (1^(st)and 3^(rd) quartile) p-value* day-180 Total IPSS score Placebo 9.1 ± 2.07.4 ± 2.0 7.6 ± 2.6 0% 8.0 (8.0-9.0) 7.0 8.0 (6.0-9.0) (6.0-8.0)Therapeutic 9.7 ± 3.1 0.63 7.6 ± 3.7 0.60 6.6 ± 3.4 0.05 −25%composition 8.0 (8.0-10.0) 7.0 6.0 (5.0-8.0) 250 mg (5.0-10.0)Therapeutic 9.4 ± 2.0 0.26 6.5 ± 2.6 0.11 5.3 ± 2.5 <0.001* −44%composition 9.0 (8.0-10.0) 7.0 5.0 (4.0-7.0) 500 mg (4.0-8.0)Voiding/obstructive Placebo 4.9 ± 1.8 5.1 ± 2.4 4.6 ± 1.8 −20% symptomsscore 5.0 (3.75-5.0) 4.0 4.0 (3.0-6.0) (4.0-6.0) Therapeutic 3.7 ± 1.70.89 3.6 ± 2.7 0.44 2.9 ± 1.7 0.18 0% composition 3.0 (3.0-5.0) 3.0 3.0(2.0-4.0) 250 mg (2.0-5.0) Therapeutic 3.8 ± 2.4 0.55 3.4 ± 2.8 0.03*2.3 ± 1.4 <0.001* −50% composition 4.0 (2.0-5.25) 3.0 2.0 (1.0-3.0) 500mg (1.0-5.0) Storage/irritative Placebo 4.2 ± 1.3 4.6 ± 1.5 4.8 ± 1.6+13% symptoms score 4.0 (3.75-5.0) 4.0 4.5 (4.0-6.0) (4.0-5.75)Therapeutic 3.7 ± 1.4 0.27 3.8 ± 1.8 0.72 3.6 ± 2.0 0.13 −13%composition 4.0 (3.0-4.25) 4.0 3.5 (2.0-5.0) 250 mg (2.0-4.75)Therapeutic 3.7 ± 1.4 0.18 3.3 ± 1.5 0.61 3.0 ± 2.0 0.018* −25%composition 4.0 (3.0-5.0) 3.0 3.0 (2.0-4.0) 500 mg (2.0-4.0) Quality ofLife Placebo 2.4 ± 0.9 2.1 ± 0.8 1.9 ± 0.7 0% 2.0 (2.0-3.0) 2.0 2.0(2.0-2.0) (1.75-2.0) Therapeutic 2.4 ± 0.8 0.64 2.2 ± 0.9 0.52 2.0 ± 0.70.71 0% composition 2.0 (2.0-3.0) 2.0 2.0 (2.0-2.0) 250 mg (2.0-3.0)Therapeutic 2.1 ± 0.7 0.03 2.0 ± 0.7 0.72 1.9 ± 0.5 0.63 0% composition2.0 (2.0-2.0) 2.0 2.0 (2.0-2.0) 500 mg (2.0-2.0) *Denotes a significantdifference versus placebo using the Mann-Whitney U test at eachindividual time point (p < 0.05).

In Table 8, results represent the mean±Standard Error of the Mean (SEM).

TABLE 8 Day-90 Day-0 IPSS Change Day-180 IPSS Change IPSS Total versusbaseline versus baseline Group Mean ± SEM Mean ± SEM Mean ± SEM Placebo9.1 ± 0.3 −1.71 ± 0.36 −1.54 ± 0.33 Therapeutic 9.7 ± 0.5 −2.16 ± 0.38−3.09 ± 0.46* Composition 250 mg Therapeutic 9.4 ± 0.3 −2.89 ± 0.36*−4.08 ± 0.31* Composition 500 mg *Denotes a significant differenceversus placebo using the Mann-Whitney U test at each individual timepoint (p < 0.05).

The mean difference in IPSS between baseline, 3-month and 6-month forthe three groups is plotted in FIG. 2. Data points represent the meandifference±the standard error of the mean. Asterisk indicates p<0.05versus placebo based on analysis of covariance at the end of treatment.At 6-month, mean difference and corresponding 95% confidence interval(CI) were −1.5 (−2.2, −0.89) for the placebo group, −3.1 (−4.0, −2.2)for the therapeutic composition 250 mg group, and −4.1 (−4.7, −3.5) forthe therapeutic composition 500 mg group.

Analysis of covariance for IPSS at 6-month with baseline IPSS entered asa covariate showed a significant dose effect (t119=−4.8, p<0.0001) and asignificant effect of baseline score (t119=8.3, p<0.0001).

IPSS voiding subscore was significantly decreased when compared toplacebo in the therapeutic composition 500 mg group 3 months afterinitiation of the treatment and up to 6 months (p=0.03 and p<0.001,respectively), while the IPSS storage subscore was significantlydecreased versus placebo only at the end of the study (p=0.018). Therewas no statistically significant improvement in the IPSS storagesubscore in the group treated with therapeutic composition at 250 mg.There was no change in the QoL questionnaire for both doses.

There was a significant improvement in all uroflowmetry parametersstudied versus baseline at the end of the intervention with therapeuticcomposition at the 500 mg dose (p<0.05). Table 9 shows LUT functionmeasurements at baseline (Day-0) and 6 months (Day-180) after placebo,therapeutic composition at 250 mg or 500 mg. Results in Table 9 arepresented as mean±SD.

TABLE 9 Q_(max), mL/s Q_(ave), mL/s PVR, mL Vol, mL Placebo Day-0 22.0 ±7.8 14.3 ± 5.2 15.0 ± 19.2 408.5 ± 117.9 Day-180 21.9 ± 8.6 14.2 ± 5.114.4 ± 18.3 364.3 ± 112.5 p-value 0.424 0.722 0.956 0.004 TherapeuticDay-0 20.5 ± 7.1 12.5 ± 4.6 15.9 ± 23.2 339.9 ± 114.4 compositionDay-180 21.4 ± 6.7 13.2 ± 4.0 13.6 ± 18.1 368.6 ± 104.6 250 mg p-value0.191 0.107 0.588 0.279 Therapeutic Day-0 19.5 ± 7.5 12.5 ± 5.4 17.8 ±21.0 339.0 ± 118.9 composition Day-180 21.7 ± 8.9 13.8 ± 5.7 9.9 ± 13.6393.0 ± 134.0 500 mg p-value  0.018*  0.040*  0.027*  0.014* *Denotes asignificant difference versus baseline using the Wilcoxon Matched-Pairstest.

Further analysis was performed to evaluate whether there was adose-response effect at the end of the intervention and whether the datacould be adjusted based on baseline data. A large proportion ofparticipants reported no measurable PVR (17/41 for 0 mg dose; 22/43 forthe 250 mg dose; and 19/38 for the 500 mg dose). While existence of anon-zero PVR was nominally lower in the 250 and 500 mg groups, no doseeffect was found. Given the existence of residual volume however, thatvolume was significantly associated with dose (z=−3.0, p=0.003). Thismodel indicates that among participants with non-zero PVR, residualurine volume would be expected to decrease by a factor of 0.09 (95% CI:0.03-0.14) for every 250 mg increase in dose, for a given baseline PVRand IPSS.

Clinical hematology parameters, which were within normal range at thebeginning and at the end of the study, thereby demonstrating the safetyof the intervention product.

This double-blind, randomized, placebo controlled study demonstrated theefficacy and the safety of the daily intake of the therapeuticcomposition at 250 or 500 mg in men with LUTS for 6 months. At the endof the study, the decrease in IPSS score was significant and dosedependent (−3.1 and −4.1 in the 250 mg and 500 mg groups, p=0.05 andp<0.001, respectively) versus the placebo group (−1.5), while no sideeffects were observed.

1. A therapeutic composition comprising dried cranberry powder and driedcranberry seeds.
 2. A therapeutic composition comprising dried cranberrypowder and cranberry seed meal.
 3. The composition of claim 1 or 2,wherein the cranberry is Vaccinium macrocarpon.
 4. The composition ofclaim 1 or 2, wherein the cranberry is Vaccinium microcarpon.
 5. Thecomposition of claim 1 or 2, wherein the cranberry is Vacciniumoxycoccus.
 6. The composition of claim 1, wherein the cranberry seedsare present in an amount of about 5% to about 50% by weight of the driedcranberry powder.
 7. The composition of claim 6, wherein the cranberryseeds are present in an amount of about 15% to about 25% by weight ofthe dried cranberry powder.
 8. The composition of claim 7, wherein thecranberry seeds are present in an amount of about 20% by weight of thedried cranberry powder.
 9. The composition of claim 2, wherein thecranberry seed meal is present in an amount of about 5% to about 50% byweight of the dried cranberry powder.
 10. The composition of claim 9,wherein the cranberry seed meal is present in an amount of about 15% toabout 25% by weight of the dried cranberry powder.
 11. The compositionof claim 10, wherein the cranberry seed meal is present in an amount ofabout 20% by weight of the dried cranberry powder.
 12. The compositionof claim 1 or 2, wherein the composition comprises less than about 12%of organic acids by weight.
 13. The composition of claim 1 or 2, whereinthe composition comprises less than 10% of organic acids by weight. 14.The composition of claim 1 or 2, wherein the composition comprises about5% to about 8% of organic acids by weight.
 15. The composition of claim1 or 2, wherein the composition comprises less than about 15% of sugarsby weight.
 16. The composition of claim 15, wherein the compositioncomprises less than about 12% of sugars by weight.
 17. The compositionof claim 1 or 2, wherein the composition comprises from about 1% toabout 5% of quinic acid by weight.
 18. The composition of claim 17,wherein the composition comprises from about 2.2% to about 3.2% ofquinic acid by weight.
 19. The composition of claim 1 or 2, wherein thecomposition comprises from about 0.4% to about 4% of malic acid byweight.
 20. The composition of claim 19, wherein the compositioncomprises from about 0.8% to about 1.8% of malic acid by weight.
 21. Thecomposition of claim 1 or 2, wherein the composition comprises fromabout 1% to about 5% of citric acid by weight.
 22. The composition ofclaim 21, wherein the composition comprises from about 1.8% to about3.2% of citric acid by weight.
 23. The composition of claim 1 or 2,wherein the composition comprises: 0.5% to 5.0% proanthocyanidins, 0.05%to 1.5% quercetin, 0.001% to 0.1% quercetin-3-glucoside, 0.001% to 0.1%quercetin-3-rhamnoside, 0.001% to 0.1% quercetin-3-xyloside, 0.001% to0.1% quercetin-3-arabinoside, 0.001% to 0.5% myricetin, 0.001% to 0.1%peonidin-3-galactoside, 0.001% to 0.1% peonidin-3-glucoside, 0.001% to0.1% peonidin-3-arabinoside, 0.001% to 0.1% cyanidin-3-glucoside, 0.001%to 0.1% cyanidin-3-galactoside, 0.001% to 0.1% cyanidin-3-arabinoside,0.001% to 0.1% protocatechuic acid 0.001% to 0.1% p-coumaric acid,0.001% to 0.1% caffeoyl-glucoside, 0.001% to 0.1% coumaroyl-glucoside,0.001% to 0.1% cafeic acid, 0.001% to 0.1% chlorogenic acid or 0.01 to1.5% ursolic acid by weight.
 24. The composition of claim 23, whereinthe composition comprises: 1.0% to 1.2% proanthocyanidins, 0.16% to0.20% quercetin, 0.07% to 0.09% quercetin-3-glucoside, 0.03% to 0.04%quercetin-3-rhamnoside, 0.019% to 0.025% quercetin-3-xyloside, 0.025% to0.035% quercetin-3-arabinoside, 0.010% to 0.014% myricetin, 0.022% to0.030% peonidin-3-galactoside, 0.0025% to 0.0035% peonidin-3-glucoside,0.010% to 0.020% peonidin-3-arabinoside, 0.0005% to 0.0015%cyanidin-3-glucoside, 0.015% to 0.030% cyanidin-3-galactoside, 0.010% to0.025% cyanidin-3-arabinoside, 0.019% to 0.025% protocatechuic acid,0.04% to 0.06% p-coumaric acid, 0.015% to 0.025% caffeoyl-glucoside,0.005% to 0.015% coumaroyl-glucoside, 0.010% to 0.015% cafeic acid or0.030% to 0.04% chlorogenic acid by weight.
 25. The composition of claim24, wherein the composition comprises about: 1.1% proanthocyanidins,0.18% quercetin, 0.083% quercetin-3-glucoside, 0.034%quercetin-3-rhamnoside, 0.022% quercetin-3-xyloside, 0.030%quercetin-3-arabinoside, 0.012% myricetin, 0.027%peonidin-3-galactoside, 0.003% peonidin-3-glucoside, 0.014%peonidin-3-arabinoside, 0.001% cyanidin-3-glucoside, 0.022%cyanidin-3-galactoside, 0.018% cyanidin-3-arabinoside, 0.022%protocatechuic acid, 0.052% p-coumaric acid, 0.021% caffeoyl-glucoside,0.011% coumaroyl-glucoside, 0.014% cafeic acid, 0.034% chlorogenic acidor 0.92% ursolic acid by weight.
 26. The composition of claim 1 or 2,wherein the composition comprises 1 to 100 μg lariciresinol, 1 to 100 μgsecoisolariciresinol or 1 to 100 μg/pinoresinol per 100 g of thecomposition by weight.
 27. The composition of claim 26, wherein thecomposition comprises about 51 μg lariciresinol, about 12 μgsecoisolariciresinol or about 78 μg/pinoresinol per 100 g of thecomposition by weight.
 28. A solid oral dosage comprising thetherapeutic composition of claim 1 or
 2. 29. The solid oral dosage ofclaim 28 which is a tablet.
 30. The solid oral dosage of claim 28 whichis a capsule.
 31. The solid oral dosage of claim 28 which is a softgel.32. The solid oral dosage of claim 30 comprising 100 mg to 500 mg of thetherapeutic composition.
 33. The solid oral dosage of claim 32comprising 250 mg of the therapeutic composition.
 34. The solid oraldosage of claim 32 comprising 500 mg of the therapeutic composition. 35.A method for alleviating lower urinary tract symptoms (LUTS) in asubject comprising administering to the subject in need thereof aneffective amount of the composition of claim 1 or
 2. 36. A method foralleviating the symptoms of benign prostatic hyperplasia (BPH) in asubject comprising administering to the subject in need thereof aneffective amount of the composition of claim 1 or
 2. 37. A method fortreating erectile dysfunction (ED) in a subject comprising administeringto the subject in need thereof an effective amount of the composition ofclaim 1 or
 2. 38. A method for treating urinary incontinence in asubject comprising administering to the subject in need thereof aneffective amount of the composition of claim 1 or
 2. 39. A method fortreating overactive bladder (OAB) in a subject comprising administeringto the subject in need thereof an effective amount of the composition ofclaim 1 or
 2. 40. A method for treating bladder obstruction,interstitial cystitis, underactive bladder, prostatitis, bladder andprostate inflammation, prostate fibrosis or pelvic pain in a subjectcomprising administering to the subject in need thereof an effectiveamount of the composition of claim 1 or
 2. 41-43. (canceled)